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Highly specific gene silencing in a monocot species by artificial microRNAs derived from chimeric MIRNA precursors

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Highly specific gene silencing in a monocot species by artificial microRNAs derived from chimeric MIRNA precursors

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dc.contributor.author CARBONELL, ALBERTO es_ES
dc.contributor.author Fahlgren, Noah es_ES
dc.contributor.author Mitchell, Skyler es_ES
dc.contributor.author Cox, Kevin L., Jr. es_ES
dc.contributor.author Reilly, Kevin C. es_ES
dc.contributor.author Mockler, Todd C. es_ES
dc.contributor.author Carrington, James C. es_ES
dc.date.accessioned 2021-02-13T04:32:04Z
dc.date.available 2021-02-13T04:32:04Z
dc.date.issued 2015-06 es_ES
dc.identifier.issn 0960-7412 es_ES
dc.identifier.uri http://hdl.handle.net/10251/161209
dc.description.abstract [EN] Artificial microRNAs (amiRNAs) are used for selective gene silencing in plants. However, current methods to produce amiRNA constructs for silencing transcripts in monocot species are not suitable for simple, cost-effective and large-scale synthesis. Here, a series of expression vectors based on Oryza sativa MIR390 (OsMIR390) precursor was developed for high-throughput cloning and high expression of amiRNAs in monocots. Four different amiRNA sequences designed to target specifically endogenous genes and expressed from OsMIR390-based vectors were validated in transgenic Brachypodium distachyon plants. Surprisingly, amiRNAs accumulated to higher levels and were processed more accurately when expressed from chimeric OsMIR390-based precursors that include distal stem-loop sequences from Arabidopsis thaliana MIR390a (AtMIR390a). In all cases, transgenic plants displayed the predicted phenotypes induced by target gene repression, and accumulated high levels of amiRNAs and low levels of the corresponding target transcripts. Genome-wide transcriptome profiling combined with 5¿-RLM-RACE analysis in transgenic plants confirmed that amiRNAs were highly specific. es_ES
dc.description.sponsorship We thank Goretti Nguyen, Robyn Stevens, Jacob Mreen, Fangfang Ma and Madison Schniers for invaluable technical assistance, and Zacchery R. Smith for his initial contribution to develop the pH7WG2B-OsMIR390-B/c vector. Noah Fahlgren was supported by a USDA AFRI NIFA Postdoctoral Fellowship (MOW-2012-01361). This work was supported by grants from the National Science Foundation (MCB-1231726, MCB-1330562) and National Institutes of Health (AI043288) to James C. Carrington, and from the Department of Energy (DOE DE-SC0006627) to Todd C. Mockler. es_ES
dc.language Inglés es_ES
dc.publisher Blackwell Publishing es_ES
dc.relation.ispartof The Plant Journal es_ES
dc.rights Reconocimiento (by) es_ES
dc.subject RNA silencing es_ES
dc.subject Artificial microRNA es_ES
dc.subject MIRNA precursor es_ES
dc.subject Brachypodium distachyon es_ES
dc.subject Monocot es_ES
dc.subject Arabidopsis thaliana es_ES
dc.subject Technical advance es_ES
dc.title Highly specific gene silencing in a monocot species by artificial microRNAs derived from chimeric MIRNA precursors es_ES
dc.type Artículo es_ES
dc.identifier.doi 10.1111/tpj.12835 es_ES
dc.relation.projectID info:eu-repo/grantAgreement/NSF//1231726/US/Function of Arabidopsis Small RNA-ARGONAUTE Complexes/ es_ES
dc.relation.projectID info:eu-repo/grantAgreement/NIH//AI043288/ es_ES
dc.relation.projectID info:eu-repo/grantAgreement/NSF//1330562/US/Integration of developmental signals by plant ARGONAUTES/ es_ES
dc.relation.projectID info:eu-repo/grantAgreement/NIFA//MOW-2012-01361/ es_ES
dc.relation.projectID info:eu-repo/grantAgreement/DOE//DE-SC0006627/ es_ES
dc.rights.accessRights Abierto es_ES
dc.contributor.affiliation Universitat Politècnica de València. Instituto Universitario Mixto de Biología Molecular y Celular de Plantas - Institut Universitari Mixt de Biologia Molecular i Cel·lular de Plantes es_ES
dc.description.bibliographicCitation Carbonell, A.; Fahlgren, N.; Mitchell, S.; Cox, KLJ.; Reilly, KC.; Mockler, TC.; Carrington, JC. (2015). Highly specific gene silencing in a monocot species by artificial microRNAs derived from chimeric MIRNA precursors. The Plant Journal. 82(6):1061-1075. https://doi.org/10.1111/tpj.12835 es_ES
dc.description.accrualMethod S es_ES
dc.relation.publisherversion https://doi.org/10.1111/tpj.12835 es_ES
dc.description.upvformatpinicio 1061 es_ES
dc.description.upvformatpfin 1075 es_ES
dc.type.version info:eu-repo/semantics/publishedVersion es_ES
dc.description.volume 82 es_ES
dc.description.issue 6 es_ES
dc.identifier.pmid 25809382 es_ES
dc.identifier.pmcid PMC4464980 es_ES
dc.relation.pasarela S\378028 es_ES
dc.contributor.funder U.S. Department of Energy es_ES
dc.contributor.funder National Science Foundation, EEUU es_ES
dc.contributor.funder National Institutes of Health, EEUU es_ES
dc.contributor.funder National Institute of Food and Agriculture, EEUU es_ES
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